Biotechnology and Recombinant DNA

Question 1

What is Gene Cloning?

Answer 1

creating the exact copy or clone of a particular gene or DNA sequence.

Question 2

How many plasmids do prokaryotes have?

Answer 2

Multiple

Question 3

Type II restriction enzymes cut where?

Answer 3

Cut DNA at a defined position close to or inside that recognized sequence.

Question 4

What bases do restriction enzymes frequently recognize?

Answer 4

4- , 6- , or 8- bases.

Question 5

Blunt ends can match flush, what of sticky ends?

Answer 5

They have to complement each other to match.

Question 6

Why are restriction enzymes used?

Answer 6

To protect the bacterial DNA from foreign DNA

Question 7

What are Vectors?

Answer 7

carrying desired genes into a new cell

Question 8

Colony Selection is better, why?

Answer 8

You will always get one definite outcome.

Question 9

Shuttle vectors?

Answer 9

vectors that can be selected in at least 2 different species

Question 10

Religation vs insertion. Which is much more than likely to occur?

Answer 10

Religation.

Question 11

How do you detect a product in PCR?

Answer 11

SYBR and TaqMan

Question 12

rtPCR

Answer 12

Reverse transcription PCR; amplifies RNA.

Question 13

rtPCR used what primers?

Answer 13

Poly AAA

Question 14

Transformation is?

Answer 14

DNA from the environment and grow infinitely

Question 15

Transfection is what?

Answer 15

DNA from the environment but added to mammalian cells.

Question 16

What are the purpose of Calcium chloride and Lithium Chloride for transformation?

Answer 16

Attaches to the outside of the cell, because the membrane is negatively charged. Causes a pore to appear on the surface.

Question 17

What is Partial Digestion?

Answer 17

random cuts by restriction enzymes.

Question 18

Average Gene is how much ?

Answer 18

1000 Bp

Question 19

Genomic Library Constructions

Answer 19

Partial digestion, creating clones with fragments and transforming to bacteria! You want a lot!

Question 20

cDNA is made from?

Answer 20

mRNA.

Question 21

Are all genes there in the cDNA library?

Answer 21

No, not all the genes are turned on at the same time. So only the genes that were turned on will be represented.

Question 22

Human insulin was used for what technology?

Answer 22

rDNA technology

Question 23

What approaches are used to treat genetic diseases (3)?

Answer 23

1. Gene therapy
2. Gene silencing
3. Gene editing

Question 24

Disadvantages of Gene therapy?

Answer 24

DNA could be inserted anywhere.

Question 25

What two ways can you accomplish gene silencing?

Answer 25

1. siRNA and 2. microRNA

Question 26

RISC is what?

Answer 26

RNA induced silencing complex incorporating siRNA or microRNA

Question 27

What does siRNA recognize?

Answer 27

long dsRNA, viruses, transposons. This is a defense mechanism.

Question 28

What do you need to accomplish Gene editing?

Answer 28

Cas9, Guide RNA, Piece of DNA

Question 29

How was the Human genome project accomplished?

Answer 29

Next-generation sequencing

Question 30

Herbicide “RoundUp” inhibits what?

Answer 30

Inhibits an enzyme in plants that creates aromatic amino acids.